Cephem compounds and processes for preparation thereof

ABSTRACT

7 β-[2-5 amino-1, 2, 4 thiadiazol-3yl) 2(carboxy lower alkyleneoxyimino) acetamido]-3-(3-amino-2-alkyl-1-pyrazolio) methyl 3 cephem 4 carboxylates are prepared. They are antibiotics.

The present invention relates to new cephem compounds andpharmaceutically acceptable salts thereof. More particularly, it relatesto new cephem compounds and pharmaceutically acceptable salts thereof,which have antimicrobial activities, to process for preparation thereof,to pharmaceutical composition comprising the same, and to a method fortreating infectious diseases in human being and animals.

Accordingly, one object of the present invention is to provide thecephem compounds and pharmaceutically acceptable salts thereof, whichare highly active against a number of pathogenic microorganisms.

Another object of the present invention is to provide processes for thepreparation of the cephem compounds and salts thereof.

A further object of the present invention is to provide pharmaceuticalcompositions comprising, as an active ingredient, said cephem compoundsor their pharmaceutically acceptable salts.

Still further object of the present invention is to provide a method fortreating infectious diseases caused by pathogenic microorganisms, whichcomprises administering said cephem compounds to infected human being oranimals.

The object cephem compounds of the present invention are novel and canbe represented by the following general formula [I] : ##STR1## wherein

R¹ and R⁴ are each amino or a protected amino group,

R² is carboxy(lower)alkyl or a protected carboxy(lower)alkyl,

R³ is lower alkyl, hydroxy(lower)alkyl or a protectedhydroxy(lower)alkyl and

R⁵ is hydrogen or lower alkyl.

As to the object compounds [I] , the following points are to be noted.

That is, the object compounds [I] include syn isomer, anti isomer and amixture thereof. Syn isomer means one geometrical isomer having thepartial structure represented by the following formula : ##STR2##(wherein R¹ and R² are each as defined above) and anti isomer means theother geometrical isomer having the partial structure represented by thefollowing formula : ##STR3## (wherein R¹ and R² are each as definedabove), and all of such geometrical isomers and mixture thereof areincluded within the scope of this invention.

In the present specification and claim, the partial structure of thesegeometrical isomers and mixture thereof are represented for convenientsake by the following formula : ##STR4## (wherein R¹ and R² are each asdefined above).

Another point to be noted is that the pyrazolio moiety of the compounds[I] can also exist in the tautomeric form, and such tautomericequilibrium can be represented by the following schemes. ##STR5##(wherein R³, R⁴ and R⁵ are each as defined above).

Both of the above tautomeric isomers are included within the scope ofthe present invention, and in the present specification and claim,however, the object compounds [I] are represented for the convenientsake by one expression of the pyrazolio group of the formula (A).

The cephem compounds [I] of the present invention can be prepared byprocesses as illustrated in the followings. ##STR6## wherein

R¹, R², R³, R⁴ and R5 are each as defined above,

R_(a) ² is a protected carboxy(lower)alkyl,

R_(bhu) 2 is carboxy(lower)alkyl, and

Y is a leaving group.

In the above and subsequent descriptions of this specification, suitableexamples of the various definitions are explained in detail as follows :

The term "lower" is intended to mean 1 to 6 carbon atom(s), unlessotherwise indicated.

Suitable "protective group" in the "protected amino group" may be loweralkanoyl [e.g. formyl, acetyl, propionyl, hexanoyl, pivaloyl, etc.],mono(or di or tri)halo(lower)alkanoyl [e.g. chloroacetyl,trifluoroacetyl, etc.], lower alkoxycarbonyl [e.g. methoxycarbonyl,ethoxycarbonyl, tert-butoxycarbonyl, tert-pentyloxycarbonyl,hexyloxycarbonyl, etc.], carbamoyl, aroyl [e.g. benzoyl, toluoyl,naphthoyl, etc.], ar(lower)alkanoyl [e.g. phenylacetyl, phenylpropionyl,etc.], aryloxycarbonyl [e.g. phenoxycarbonyl, naphthyloxycarbonyl,etc.], aryloxy(lower)alkanoyl [e.g. phenoxyacetyl, phenoxypropionyl,etc.], arylglyoxyloyl [e.g. phenylglyoxyloyl, naphthylglyoxyloyl, etc.],ar(lower)alkoxycarbonyl which may have suitable substituent(s) [e.g.benzyloxycarbonyl, phenethyloxycarbonyl, p-nitrobenzyloxycarbonyl,etc.], substituted or unsubstituted ar(lower)alkylidene [e.g.benzylidene, hydroxybenzylidene, etc.], ar(lower)alkyl such as mono ordi or triphenyl(lower)alkyl [e.g. benzyl, phenethyl, benzhydryl, trityl,etc.], or the like, in which the preferred one may be lower alkanoyl orcarbamoyl and the more preferred one may be C₁ -C₄ alkanoyl orcarbamoyl.

Suitable "lower alkyl" and "lower alkyl moieties"in the"caboxy(lower)alkyl", "protected carboxy(lower)-alkyl","hydroxy(lower)alkyl" and "protected hydroxy-(lower)alkyl"may be astraight or branched one such as methyl, ethyl, propyl, isopropyl,butyl, isobutyl, tert-butyl, pentyl, hexyl, or the like, in which thepreferred one may be C₁ -C₄ alkyl.

Suitable "protected carboxy" in the "protected carboxy(lower)alkyl" maybe an esterified carboxy group, or the like, and concrete examples ofthe ester moiety in said esterified carboxy group may be the ones suchas lower alkyl ester [e.g. methyl ester, ethyl ester, propyl ester,isopropyl ester, butyl ester, isobutyl ester, tert-butyl ester, pentylester, hexyl ester, 1-cyclopropylethyl ester, etc.] which may havesuitable substituent(s), for example, lower alkanoyloxy(lower)alkylester [e.g. acetoxymethyl ester, propionyloxymethyl ester,butyryloxymethyl ester, valeryloxymethyl ester, pivaloyloxymethyl ester,1-acetoxyethyl ester, 1-propionyloxyethyl ester, pivaloyloxymethylester, 2-propionyloxyethyl ester, hexanoyloxymethyl ester, etc.], loweralkanesulfonyl(lower)alkyl ester [e.g. 2-mesylethyl ester, etc.] ormono(or di or tri)halo(lower)alkyl ester [e.g. 2-iodoethyl ester,2,2,2-trichloroethyl ester, etc.]; lower alkenyl ester [e.g. vinylester, allyl ester, etc.]; lower alkynyl ester [e.g. ethynyl ester,propynyl ester, etc.]; ar(lower)alkyl ester which may have suitablesubstituent(s) [e.g. benzyl ester, 4-methoxybenzyl ester, 4-nitrobenzylester, phenethyl ester, trityl ester, benzhydryl ester,bis(methoxyphenyl)methyl ester, 3,4-dimethoxybenzyl ester,4-hydroxy-3,5-di-tert-butylbenzyl ester, etc.]; aryl ester which mayhave suitable substituent(s) [e.g. phenyl ester, 4-chlorophenyl ester,tolyl ester, 4-tert-butylphenyl ester, xylyl ester, mesityl ester,cumenyl ester, etc.]; or the like.

Suitable "protected hydroxy" in the "protected hydroxy(lower)alkyl" maybe acyloxy group or the like. Suitable "acyl moiety" in the "acyloxy"may be lower alkanoyl [e.g. formyl, acetyl, propionyl, hexanoyl,pivaloyl, etc., mono(or di or tri)halo(lower)alkanoyl [e.g.chloroacetyl, trifluoroacetyl, etc.], lower alkoxycarbonyl [e.g.methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl,tert-pentyloxycarbonyl, hexyloxycarbonyl, etc.]or the like.

Suitable "leaving group" may be halogen [e.g. chlorine, bromine, iodine,etc.], acyloxy such as sulfonyloxy [e.g. benzenesulfonyloxy, tosyloxy,mesyloxy, etc.], lower alkanoyloxy [e.g. acetyloxy, propionyloxy, etc.],or the like.

Suitable pharmaceutically acceptable salts of the object compounds [I]are conventional non-toxic salts and include a metal salt such as analkali metal salt [e.g. sodium salt, potassium salt, etc.] and analkaline earth metal salt [e.g. calcium salt, magnesium salt, etc.], anammonium salt, an organic base salt [e.g. trimethylamine salt,triethylamine salt, pyridine salt, picoline salt, dicyclohexylaminesalt, N,N'-dibenzylethylenediamine salt, etc.], an organic acid salt[e.g. formate, acetate, trifluoroacetate, maleate, tartrate,methanesulfonate, benzenesulfonate, toluenesulfonate, etc.], aninorganic acid salt [e.g. hydrochloride, hydrobromide, sulfate,phosphate, etc.], a salt with an amino acid [e.g. arginine salt,aspartic acid salt, glutamic acid salt, etc.], and the like.

Preferred embodiments of the object compound [I]are as follows.

Preferred embodiment of R¹ is amino, R² is carboxy(lower)alkyl [morepreferably carboxy-(C₁ -C₄)alkyl, most preferably carboxymethyl or1-carboxy-1-methylethyl] or an esterfied carboxy(lower)-alkyl [morepreferably lower alkoxycarbonyl(lower)-alkyl, most preferably loweralkoxycarbonyl(C₁ -C₄)-alkyl],

R³ is lower alkyl [more preferably (C₁ -C₄)alkyl, most preferablymethyl] or hydroxy(lower)alkyl [more preferably hydroxy(C₁ -C₄)alkyl,most preferably hydroxyethyl],

R⁴ is amino, lower alkanoylamino or ureido,

R⁵ is hydrogen or lower alkyl [more preferably (C_(l) -C₄)-alkyl, mostpreferably methyl].

The processes for preparing the object compounds of the presentinvention are explained in detail in the following.

Process 1

The object compound [I] or a salt thereof can be prepared by reacting acompound [II] or its reactive derivative at the amino group or a saltthereof with a compound [III] or its reactive derivative at the carboxygroup or a salt thereof.

Suitable reactive derivative at the amino group of the compound [II] mayinclude Schiff's base type imino or its tautomeric enamine type isomerformed by the reaction of the compound [II] with a carbonyl compoundsuch as aldehyde, ketone or the like; a silyl derivative formed by thereaction of the compound [II] with a silyl compound such asbis(trimethylsilyl)acetamide, mono(trimethylsilyl)acetamide [e.g.N-(trimethylsilyl)acetamide], bis(trimethylsilyl)urea or the like; aderivative formed by reaction of the compound [II]with phosphorustrichloride or phosgene, and the like.

Suitable 1salts of the compound [II] and its reactive derivative can bereferred to the ones as exemplified for the compound [I].

Suitable reactive derivative at the carboxy group of the compound [III]may include an acid halide, an acid anhydride, an activated amide, anactivated ester, and the like. Suitable examples of the reactivederivatives may be an acid chloride; an acid azide; a mixed acidanhydride with an acid such as substituted phosphoric acid [e.g.dialkylphosphoric acid, phenylphosphoric acid, diphenylphosphoric aciddibenzylphosphoric acid, halogenated phosphoric acid, etc.],dialkylphosphorous acid, sulfurous acid, thiosulfuric acid, sulfuricacid, sulfonic acid [e.g. methanesulfonic acid, etc.], aliphaticcarboxylic acid [e.g. acetic acid, propionic acid, butyric acid,isobutyric acid, pivalic acid, pentanoic acid, isopentanoic acid,2-ethylbutyric acid, trichloroacetic acid, etc.] or aromatic carboxylicacid [e.g. benzoic acid, etc.]; a symmetrical acid anhydride; anactivated amide with imidazole, 4-substituted imidazole,dimethylpyrazole, triazole or tetrazole; or an activated ester [e.g.cyanomethyl ester, methoxymethyl ester, dimethylliminomethyl [(CH₃)₂ N⁺═CH--] ester, vinyl ester, propargyl ester, p-nitrophenyl ester,2,4-dinitrophenyl ester, trichlorophenyl ester, pentachlorophenyl ester,mesylphenyl ester, phenylazophenyl ester, phenyl thioester,p-nitrophenyl thioester, p-cresyl thioester, carboxymethyl thioester,pyranyl ester, pyridyl ester, piperidyl ester, 8-quinolyl thioester,etc.], or an ester with a N-hydroxy compound [e.g.N,N-dimethylhydroxylamine, 1-hydroxy-2-(1H)-pyridone,N-hydroxysuccinimide, N-hydroxyphthalimide, 1-hydroxy-1H-benzotriazole,etc.], and the like. These reactive derivatives can optionally beselected from them according to the kind of the compound [III] to beused.

Suitable salts of the compound [III] and its reactive derivative can bereferred to the ones as exemplified for the compound [I].

The reaction is usually carried out in a conventional solvent such aswater, alcohol [e.g. methanol, ethanol, etc.], acetone, dioxane,acetonitrile, chloroform, methylene chloride, ethylene chloride,tetrahydrofuran, ethyl acetate, N,N-dimethylformamide, pyridine or anyother organic solvent which does not adversely influence the reaction.These conventional solvent may also be used in a mixture with water.

In this reaction, when the compound [III] is used in a free acid form orits salt form, the reaction is preferably carried out in the presence ofa conventional condensing agent such as N,N'-dicyclohexylcarbodiimide;N-cyclohexyl-N'-morpholinoethylcarbodiimide;N-cyclohexyl-N'-(4-diethylaminocyclohexyl)carbodiimide;N,N'-diethylcarbodiimide, N,N'-diisopropylcarbodiimide;N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide;N,N'carbonylbis-(2-methylimidazole);pentamethyleneketene-N-cyclohexylimine;diphenylketene-N-cyclohexylimine; ethoxyacetylene;1-alkoxy-1-chloroethylene; trialkyl phosphite; ethyl polyphosphate;isopropyl polyphosphate; phosphorus oxychloride (phosphoryl chloride);phosphorus trichloride; thionyl chloride; oxalyl chloride; lower alkylhaloformate [e.g. ethyl chloroformate, isopropyl chloroformate, etc.];triphenylphosphine; 2-ethyl-7-hydroxybenzisoxazolium salt;2-ethyl-5-(m-sulfophenyl)isoxazolium hydroxide intramolecular salt;1-(p-chlorobenzenesulfonyloxy)-6-chloro-1H-benzotriazole; so-calledVilsmeier reagent prepared by the reaction of N,N-dimethylformamide withthionyl chloride, phosgene, trichloromethyl chloroformate, phosphorusoxychloride, etc.; or the like.

The reaction may also be carried out in the presence of an inorganic ororganic base such as an alkali metal bicarbonate, tri(lower)alkylamine,pyridine, N-(lower)alkylmorpholine, N,N-di(lower)alkylbenzylamine, orthe like.

The reaction temperature is not critical, and the reaction is usuallycarried out under cooling to warming.

Process 2

The object compound [Ib] or a salt thereof can be prepared by subjectinga compound [Ia] or a salt thereof to elimination reaction of the carboxyprotective group.

This reaction is carried out in accordance with a conventional methodsuch as hydrolysis, reduction or the like.

The hydrolysis is preferably carried out in the presence of base or anacid including Lewis acid. Suitable base may include an inorganic baseand an organic base such as an alkali metal [e.g. sodium, potassium,etc.], an alkaline earth metal [e.g. magnesium, calcium, etc.], thehydroxide or carbonate or bicarbonate thereof, trialkylamine [e.g.trimethylamine, triethylamine, etc.], picoline,1,5-diazabicyclo[4.3.0]non-5-ene, 1,4-diazabicyclo[2.2.2]octane,1,8-diazabicyclo[5.4.0]-undec-7-ene, or the like. Suitable acid mayinclude an organic acid [e.g. formic acid, acetic acid, propionic acid,trichloroacetic acid, trifluoroacetic acid, etc.] and an inorganic acid[e.g. hydrochloric acid, hydrobromic acid, sulfuric acid, hydrogenchloride, hydrogen bromide, etc.]. The elimination using Lewis acid suchas trihaloacetic acid [e.g. trichloroacetic acid, trifluoroacetic acid,etc.] or the like is preferably carried out in the presence of cationtrapping agents [e.g. anisole, phenol, etc.].

The reaction is usually carried out in a solvent such as water, analcohol [e.g. methanol, ethanol, etc.], methylene chloride,tetrahydrofuran, a mixture thereof or any other solvent which does notadversely influence the reaction. A liquid base or acid can be also usedas the solvent. The reaction temperature is not critical and thereaction is usually carried out under cooling to warming.

The reduction method applicable for the elimination reaction may includechemical reduction and catalytic reduction.

Suitable reducing agents to be used in chemical reduction are acombination of metal [e.g. tin, zinc, iron, etc.] or metallic compound[e.g. chromium chloride, chromium acetate, etc.] and an organic orinorganic acid [e.g. formic acid, acetic acid, propionic acid,trifluoroacetic acid, p-toluenesulfonic acid, hydrochloric acid,hydrobromic acid, etc.].

Suitable catalysts to be used in catalytic reduction are conventionalones such as platinum catalysts [e.g. platinum plate, spongy platinum,platinum black, colloidal platinum, platinum oxide, platinum wire,etc.], palladium catalysts [e.g. spongy palladium, palladium black,palladium oxide, palladium on carbon, colloidal palladium, palladium onbarium sulfate, palladium on barium carbonate, etc.], nickel catalysts[e.g. reduced nickel, nickel oxide,Raney nickel, etc.], cobalt catalysts[e.g. reduced cobalt, Raney cobalt, etc.], iron catalysts [e.g. reducediron, Raney iron, etc.], copper catalysts [e.g. reduced copper, Raneycopper, Ullman copper, etc.] and the like.

The reduction is usually carried out in a conventional solvent whichdoes not adversely influence the reaction such as water, methanol,ethanol, propanol, N,N-dimethylformamide, or a mixture thereof.Additionally, in case that the abovementioned acids to be used inchemical reduction are in liquid, they can also be used as a solvent.Further, a suitable solvent to be used in catalytic reduction may be theabovementioned solvent, and other conventional solvent such as diethylether, dioxane, tetrahydrofuran, etc., or a mixture thereof.

The reaction temperature of this reduction is not critical and thereaction is usually carried out under cooling to warming.

Process 3

The object compound [I] or a salt thereof can be prepared by reacting acompound [VII] or a salt thereof with a compound [V] or a salt thereof.

Suitable salts of the compounds [V] and [VII] can be referred to theones as exemplified for the compound [I].

The present reaction may be carried out in a solvent such as water,phosphate buffer acetone, chloroform, acetonitrile, nitrobenzene,methylene chloride, ethylene chloride, formamide, N,N-dimethylformamide,methanol, ethanol, diethyl ether, tetrahydrofuran, dimethyl sulfoxide,or any other organic solvent which does not adversely affect thereaction, preferably in ones having strong polarities. Among thesolvents, hydrophilic solvents may be used in a mixture with water. Whenthe compound [V] is in liquid, it can also be used as a solvent. Thereaction is preferably conducted in the presence of a base, for example,inorganic base such as alkali metal hydroxide, alkali metal carbonate,alkali metal bicarbonate, organic base such as trialkylamine, and thelike. The reaction temperature is not critical, and the reaction isusually carried out at ambient temperature, under warming or underheating. The present reaction is preferably carried out in the presenceof alkali metal halide [e.g. sodium iodide, potassium iodide, etc.],alkali metal thiocyanate [e.g. sodium thiocyanate, potassiumthiocyanate, etc.] or the like.

The starting compound [II] is novel and can be prepared by the followingprocesses A and B. ##STR7## wherein

R³, R⁴, R⁵ and Y are each as defined above,

R⁶ is a protected amino group,

R⁷ is a protected carboxy group,

X.sup.⊖ is an anion, and

R_(a) ⁴ is a protected amino group.

Suitable "protective group" in the "protected amino group" for R⁶ can bereferred to the ones as exemplified before and the preferred one may besubstituted or unsubstituted ar(lower)alkylidene (e.g., benzylidene,hydroxybenzylidene, etc.), lower alkoxycarbonyl, (e.g., methoxycarbonyl,ethoxycarbonyl, tert-butoxycarbonyl, tert-pentyloxycarbonyl,hexyloxycarbonyl, etc.).

Suitable "protected carboxy group" for R⁷ can be referred to the ones asexemplified before and the preferred one may be ar(lower)alkoxycarbonylwhich may have suitable substituent(s) (e.g. benzhydryloxycarbonyl,etc.).

Suitable "anion" may be formate, acetate, trifluoroacetate, maleate,tartrate, methanesulfonate, benzenesulfonate, toluenesulfonate,chloride, bromide, iodide, sulfate, phosphate, or the like.

Processes A and B for the preparation of the starting compound [II] isexplained in detail in the following.

Process A - ○1

The compound [VI] or a salt thereof can be prepared by reacting thecompound [IV] or a salt thereof with the compound [V] or a salt thereof.This reaction can be carried out in a similar manner to that of theaforementioned Process 3, and therefore the reagents to be used and thereaction conditions (e.g., solvent, reaction temperature, etc.) can bereferred to those of the Process 3.

Anion X.sup.⊖ may be the one derived from a leaving group Y and may bethe other one converted therefrom by a conventional method.

Process A - ○2

The compound [II] or a salt thereof can be prepared by subjecting thecompound [VI] or a salt thereof to elimination reaction of the aminoprotective group in R⁶ and the carboxy protective group in R⁷.

This reaction is carried out in accordance with a conventional methodsuch as hydrolysis or the like.

The hydrolysis is preferably carried out in the presence of a base or anacid including Lewis acid. Suitable base may include an inorganic baseand an organic base such as an alkali metal [e.g. sodium, potassium,etc.], an alkaline earth metal [e.g. magnesium, calcium, etc.], thehydroxide or carbonate or bicarbonate thereof, trialkylamine [e.g.trimethylamine, triethylamine, etc.], picoline,1,5-diazabicyclo-[4.3.0]non-5-ene, 1,4-diazabicyclo[2.2.2]octane,1,8-diazabicyclo[5.4.0]undec-7-ene, or the like. Suitable acid mayinclude an organic acid [e.g. formic acid, acetic acid, propionic acid,trichloroacetic acid, trifluoroacetic acid, etc.] and an inorganic acid[e.g. hydrochloric acid, hydrobromic acid, sulfuric acid, hydrogenchloride, hydrogen bromide, etc.]. The elimination using Lewis acid suchas trihaloacetic acid [e.g. trichloroacetic acid, trifluoroacetic acid,etc.] or the like is preferably carried out in the presence of cationtrapping agents [e.g. anisole, phenol, etc.].

The reaction is usually carried out in a solvent such as water, analcohol [e.g. methanol, ethanol, etc.], methylene chloride,tetrahydrofuran, a mixture thereof or any other solvent which does notadversely influence the reaction. A liquid base or acid can be also usedas the solvent. The reaction temperature is not critical and thereaction is usually carried out under cooling to warming.

The present invention includes within the scope of the invention thecase that protected amino in R⁴ is transformed into amino during thisreaction.

Process B

The compound [IIb] or a salt thereof can be prepared by subjecting thecompound [IIa] to elimination reaction of the amino protective group inR_(a) ⁴. This reaction can be carried out in a simila that of theaforementioned Process A - ○2 , therefore the reagents to be used andthe reaction conditions (e.g., solvent, reaction temperature, etc.) canbe referred to those of the Process A - ○2 . The present inventionincludes within the scope of the invention the case that protectedhydroxy(lower)alkyl in R³ is transformed into hydroxy(lower)alkyl duringthis reaction.

The object compounds [I] and pharmaceutically acceptable salts thereofare novel and exhibit high antimicrobial activity, inhibiting the growthof a wide variety of pathogenic microorganisms including Gram-positiveand Gram-negative microorganisms and are useful as antimicrobial agents.

Now in order to show the utility of the object compounds [I], the testdata on MIC (minimal inhibitory concentration) of representativecompounds [I] of this invention are shown in the following.

Test method

In vitro antibacterial activity was determined by the two-foldagar-plate dilution method as described below.

One loopful of an overnight culture of each test strain inTrypticase-soy broth (10⁶ viable cells per ml) was streaked on heartinfusion agar (HI-agar) containing graded concentrations ofrepresentative test compound, and the minimal inhibitory concentration(MIC) was expressed in terms of μg/ml after incubation at 37° C. for 20hours.

Test compounds

(1)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer) (hereinafter referred to as Compound A).

(2)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-(3-formamido-2-methyl-1-pyrazolio)-methyl-3-cephem-4-carboxylate(syn isomer) (hereinafter referred to as Compound B).

(3)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer) (hereinafter referred to as Compound C).

Test results

    ______________________________________                                                   MIC (μg/ml)                                                     Test       Test compounds                                                     strain     A             B      C                                             ______________________________________                                        P. vulgaris                                                                              0.20          0.20   0.10                                          49                                                                            ______________________________________                                    

For therapeutic administration, the object compounds [I] andpharmaceutically acceptable salts thereof of the present invention areused in the form of conventional pharmaceutical preparation whichcontains said compound as an active ingredient, in admixture withpharmaceutically acceptable carriers such as an organic or inorganicsolid or liquid excipient which is suitable for oral, parenteral andexternal administration. The pharmaceutical preparations may be in solidform such as tablet, granule, powder, capsule, or liquid form such assolution, suspension, syrup, emulsion, lemonade and the like.

If needed, there may be included in the above preparations auxiliarysubstances, stabilizing agents, wetting agents and other commonly usedadditives such as lactose, citric acid, tartaric acid, stearic acid,magnesium stearate, terra alba, sucrose, corn starch, talc, gelatin,agar, pectin, peanut oil, olive oil, cacao butter, ethylene glycol, andthe like.

While the dosage of the compound [I] may vary from and also depend uponthe age, conditions of the patient, a kind of diseases, a kind of thecompound [I] to be applied, etc. In general, amounts between 1 mg andabout 4,000 mg or even more per day may be administered to a patient. Anaverage single dose of about 50 mg, 100 mg, 250 mg, 500 mg, 1000 mg,2000 mg of the object compounds [I] of the present invention may be usedin treating diseases infected by pathogenic microorganisms.

The following Preparations and Examples are given for the purpose ofillustrating the present invention in more detail.

PREPARATION 1

A mixture of acetic anhydride (38.86 ml) and formic acid (15.54 ml) wasstirred at 45° C. for 45 minutes. To this mixture was added5-amino-1-methylpyrazole (10 g) under ice-cooling, and the reactionmixture was stirred at the same temperature for 10 minutes. Theresultant mixture was poured into a mixture of water and ethyl acetate,and the resultant solution was adjusted to pH 8 with potassiumcarbonate. The organic layer was separated, and the aqueous layer wasextracted with ethyl acetate six times. The organic layers werecombined, dried over magnesium sulfate, and evaporated in vacuo to give5-formamido-1-methylpyrazole (12.88 g).

mp:71-73° C.

IR (Nujol) : 3300, 3200, 1705, 1590 cm⁻¹

NMR (CDCl₃, δ) : 3.69 and 3.74 (3H, each s),

6.04 and 6.23 (1H, each d, J=3Hz),

7.34 (1H, s), 8.21 (1H, s)

PREPARATION 2

The following compounds were obtained according to a similar manner tothat of Preparation 1.

(1) 4-Formamido-1-methylpyrazole

mp: 44-45° C.

IR (Nujol) : 3250, 1665, 1585 cm⁻¹

NMR (CDCl₃, δ):3.83 (3H, s), 7.33 (1H, s), 7.83 (1H, s), 8.17 (1H, s)

(2) 5-Formamido-1,4-dimethylpyrazole

IR (Nujol) : 3200, 1665, 1585 cm⁻¹

NMR (CDCl₃, δ) : 1.90 and 1.98 (3H, each s), 3.64 and 3.72 (3H, each s),7.29 and 7.31 (1H, each s), 8.10 (1H, broad s), 8.33 and 9.03 (1H, eachs).

PREPARATION 3

To a mixture of benzhydryl7δ-tert-butoxycarbonylamino-3-chloromethyl-3-cephem-4-carboxylate (15 g)and sodium iodide (4.37 g) in acetone (15 ml) was added5-fomamido-1-methylpyrazole (15 g) at ambient temperature. After beingstirred for 40 hours at the same temperature, the mixture was pouredinto a mixture of water and ethyl acetate. The organic layer wasseparated and washed with water, aqueous sodium chloride solution, anddried over magnesium sulfate.

The solution was evaporated in vacuo to give benzhydryl7β-tert-butoxycarbonylamino-3-(3-formamido-2-methyl-1-pyrazolio)methyl-3-cephem.4-carboxylateiodide (20.95 g).

IR (Nujol) : 1780, 1710, 1580 cm⁻¹

NMR (DMSO-d₆, δ) : 1.40 (9H, s), 3.41 (2H, broad s), 3.65 (3H, s), 5.12(1H, d, J=5Hz), 5.36 (2H, broad s), 5.57 (1H, dd, J=8Hz and 5Hz), 6.88(1H, s), 6.89 (1H, m), 7.10-7.48 (10H, m), 7.83 (1H, d, J=8Hz), 8.24(1H, d, J=3Hz), 8.45 (1H, s)

PREPARATION 4

The following compounds were obtained according to a similar manner tothat of Preparation 3.

(1) Benzhydryl7β-tert-butoxycarbonylamino-3-(4-formamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylateiodide

IR (Nujol) : 1785, 1720, 1605 cm⁻¹

NMR (DMSO-d₆, δ) : 1.39 (9H, s), 3.42 (2H, broad s), 3.77 (3H, s), 5.11(1H, d, J=5Hz), 5.41 (2H, broad s), 5.60 (1H, dd, J=8Hz and 5Hz), 6.89(1H, s), 7.18-7.52 (10H, m), 7.96 (1H, d, J=8Hz), 8.25 (1H, s), 8.51(1H, s), 8.57 (1H, s).

(2) Benzhydryl7β-tert-butoxycarbonylamino-3-(3-formamido-2,4-dimethyl-1-pyrazolio)methyl-3-cephem-4-carboxylateiodide.

IR (Nujol) : 3300, 1780, 1705 cm⁻¹

NMR (DMSO-d₆, δ) : 1.42 (9H, s), 1.98 (3H, s), 3.45 (2H, broad s), 3.63(3H, s), 5.19 (1H, d, J=5Hz), 5.40 (2H, broad s), 5.61 (1H, dd, J=5Hzand 8Hz), 6.95 (1H, s), 7.21-7.58 (10H, m), 8.00 (1H, d, J=8Hz), 8.21(1H, s), 8.43 (1H, s)

PREPARATION 5

To a solution of benzhydryl7β-tert-butoxycarbonylamino-3-(3-formamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate iodide (20.9 g) and anisole (20ml) in methylene chloride (40 ml) was added dropwise trifluoroaceticacid (40 ml) under ice-cooling. After being stirred for 1.5 hours atambient temperature, the mixture was poured into a mixture ofdiisopropyl ether (300 ml) and ethyl acetate (300 ml). The resultantprecipitate was collected by filtration to give di(trifluoroacetic acid)salt of7β-amino-3-(3-formamido-2-methyl-1-pyrazolio)-methyl-3-cephem-4-carboxylate(16.20 g).

IR (Nujol) : 3350, 1770, 1660 cm⁻¹

NMR (DMSO-d₆, δ) : 3.45 (2H, s), 3.87 (3H, s), 5.18 (2H, s), 5.47 (2H,s), 6.95 (1H, d, J=3Hz), 8.33 (1H, d, J=3Hz), 8.47 (1H, s).

PREPARATION 6

The following compounds were obtained according to a similar manner tothat of Preparation 5.

(1) Di(trifluoroacetic acid)salt of7β-amino-3-(4-formamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate.

IR (Nujol) : 3400, 1780, 1660, 1605 cm⁻¹

NMR (DMSO-d₆, δ) : 3.51 (2H, broad s), 4.06 (3H, s), 5.23 (2H, s), 5.55(2H, broad s), 8.30 (1H, s), 8.61 (1H, s), 8.67 (1H, s).

(2) Di(trifluoroacetic acid)salt of7β-amino-3-(3-formamido-2,4-dimethyl-1-pyrazolio)methyl-3-cephem-4-carboxylate.

NMR (DMSO- d₆, δ) : 2.01 (3H, s), 3.48 (2H, broad s), 3.83 (3H, s), 5.24(2H, s), 5.50 (2H, broad s), 8.26 (1H, s), 8.41 (1H, s).

PREPARATION 7

Concentrated hydrochloric acid (0.353 ml) was added to a mixture ofdi(trifluoroacetic acid)salt of7β-amino-3-(3-formamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(0.565 g) in tetrahydrofuran (3 ml) and methanol (3 ml) at ambienttemperature. After being stirred at the same temperature for 12 hours,the mixture was added dropwise to ethyl acetate (100 ml). The resultantprecipitate was collected by filtration to give7β-amino-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylatetrihydrochloride (292 mg).

NMR (DMSO-d₆, δ) : 3.31 and 3.56 (2H, ABq, J=18 Hz), 3.67 (3H, s), 5.20(2H, broad s), 5.29 (2H, broad s), 5.87 (1H, d, J=3Hz), 8.12 (1H, d,J=3Hz)

PREPARATION 8

7β-Amino-3-(3-amino-2,4-dimethyl-1-pyrazolio)methyl-3-cephem-4-carboxylatetrihydrochloride was obtained according to a similar manner to that ofPreparation 7.

IR (Nujol) : 3350, 1780, 1640 cm⁻¹

NMR (DMSO-d₆, δ) : 1.97 (3H, s), 3.49 (2H, s), 3.74 (3H, s), 5.27 (4H,s), 8.00 (1H, s).

EXAMPLE 1

To a solution of di(trifluoroacetic acid)salt of7β-amino-3-(4-formamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(4.0 g) and N-(trimethylsilyl)acetamide (9.29 g) in tetrahydrofuran (60ml) was added methanesulfonyl(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetate (2.07g) under ice-cooling. The mixture was stirred for 1 hour at ambienttemperature and then the resulting mixture was added dropwise todiisopropyl ether. Thus produced precipitate was collected byfiltration, dissolved in water and then subjected to columnchromatography on macroporous non-ionic adsorption resin, Diaion HP-20(Trademark: manufactured by Mitsubishi Chemical Industries). The desiredproduct was eluted with 10% aqueous isopropyl alcohol solution, and thenlyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-(4-formamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carb #oxylate (synisomer 0.714 g).

IR (Nujol) : 3300, 1765, 1665, 1605 cm⁻¹

NMR (D₂ O, δ) : 3.18 and 3.51 (2H, ABq, J=18Hz), 4.09 (3H, s), 4.63 (2H,s), 5.22 and 5.46 (2H, ABq, J=15Hz), 5.23 (1H, d, J=5Hz), 5.86 (1H, d,J=5Hz), 8.25 (1H, s), 8.36 (1H, s), 8.43 (1H, s)

EXAMPLE 2

(1)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-(3-formamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer) was obtained according to a similar manner to that ofExample 1.

IR (Nujol) : 3300, 1760, 1660, 1580 cm⁻¹

NMR (D₂ O, δ) : 3.13 and 3.44 (2H, ABq, J=18Hz), 3.91 (3H, s), 4.67 (2H,s), 5.21 (1H, d, J=5Hz), 5.22 and 5.43 (2H, ABq, J=15Hz), 5.85 (1H, d,J=5Hz), 6.82 and 6.92 (1H, each d, J=3Hz), 8.15 (1H, d, J=3Hz), 8.40(1H, s).

(2)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-(3-formamido-2,4-dimethyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer) was obtained according to a similar manner to that ofExample 1.

IR (Nujol) : 3300, 1765, 1660, 1600 cm⁻¹

NMR (D₂ O, δ) : 2.03 (3H, s), 3.17 and 3.48 (2H, ABq, J=18Hz), 3.83 (3H,s), 4.70 (2H, s), 5.18 and 5.43 (2H, ABq, J=15Hz), 5.24 (1H, d, J=5Hz),5.87 (1H, d, J=5Hz), 8.05 (1H, s), 8.36 (1H, s)

EXAMPLE 3

To a solution of7β-amino-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylatetrihydrochloride (6.96 g) and N-(trimethylsilyl)acetamide (21.8 g) intetrahydrofuran (150 ml) was added methanesulfonyl(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetate (4.86g) at ambient temperature. After being stirred for 30 minutes at thesame temperature, the mixture was poured into diethyl ether (2 l), andthe resultant precipitate was collected by filtration. The precipitatewas dissolved in water, and the solution was adjusted to pH 2.0 withaqueous sodium bicarbonate solution. The solution was subjected tocolumn chromatography on macroporous non-ionic adsorption resin, DiaionHP-20. The desired product was eluted with 5% isopropyl alcohol solutionand lyophilized to give7β[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)(5.20 g).

IR (Nujol) : 3300, 1760, 1660, 1590 cm⁻¹

NMR (D₂ O, 6) : 3.06 and 3.33 (2H, ABq, J=18Hz), 3.63 (3H, s), 4.60 (2H,s), 4.93 and 5.21 (2H, ABq, J=15Hz), 5.16 (1H, d, J=5Hz), 5.83 (1H, d,J=5Hz), 5.88 (1H, d, J=3Hz), 7.78 (1H, d, J=3Hz).

EXAMPLE 4

To a solution of7β-amino-3-(3-amino-2,4-dimethyl-1-pyrazolio)methyl-3-cephem-4-carboxylatetrihydrochloride (1.0 g) and N-(trimethylsilyl)-acetamide (3.03 g) intetrahydrofuran (20 ml) was added methanesulfonyl(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetate (0.68g) at ambient temperature. After being stirred for 1 hour at the sametemperature, the mixture was poured into diethyl ether (300 ml) and theresultant precipitate was collected by filtration. The precipitate wasdissolved in water and the solution was adjusted to pH 2.0 with aqueoussodium bicarbonate solution.

The solution was subjected to a column chromatography on macroporousnon-ionic adsorption resin, Diaion HP-20. The desired product was elutedwith 5% isopropyl alcohol aqueous solution and lyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-(3-amino-2,4-dimethyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)(0.165 g).

IR (Nujol) : 3350, 1770, 1660, 1600 cm⁻¹

NMR (D₂ O, δ) 1.93 (3H, s), 3.06 and 3.30 (2H, ABq, J=18Hz), 3.64 (3H,s), 4.67 (2H, s), 4.88 and 5.19 (2H, ABq, J=15Hz), 5.18 (1H, d, J=5Hz),5.84 (1H, d, J=5Hz), 7.66 (1H, s).

PREPARATION 9

To a solution of phosphorus pentachloride (11.11 g) in methylenechloride (167.8 ml) was added(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-1-methylethoxyimino)aceticacid (16.78 g) at -20° C. The resultant mixture was stirred at -20 to-10° C. for 1.5 hours, and to the mixture was added dropwise diisopropylether (671.2 ml) at -20 to -10° C. The mixture was stirred underice-cooling for 1 hour and the resultant precipitate was collected byfiltration to give(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-1-methylethoxyimino)acetylchloride hydrochloride (14.8 g).

IR (Nujol) : 3430, 3270, 3130, 1815, 1750, 1725, 1640 cm⁻¹

EXAMPLE 5

To a solution of7β-amino-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylatetrihydrochloride (2 g) and N-(trimethylsilyl)acetamide (6.28 g) intetrahydrofuran (40 ml) was added(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-1-methyl-ethoxyimino)acetylchloride hydrochloride (1.84 g) under ice-cooling. After being stirredfor 1 hour, the reaction mixture was added dropwise to ethyl ether (300ml), and the resulting precipitate was collected by filtration to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-1-methylethoxyimino)-acetamido]-3-(3-amino-2-methyl-1pyrazolio)methyl-3-cephem-4#-carboxylate trihydrochloride (syn isomer) (3.4 g).

IR (Nu]ol) : 3300, 1780, 1720, 1650 cm⁻¹

NMR (D₂ O, δ) : 1.45 (9H, s), 1.57 (6H, s), 3.09 and 3.37 (2H, ABq,J=18Hz), 3.67 (3H, s), 4.98 and 5.27 (2H, ABq, J=15Hz), 5.21 (1H, d,J=5Hz), 5.86 (1H, d, J=5Hz), 5.92 (1H, d, J=3Hz), 7.85 (1H, d, J=3Hz)

EXAMPLE 6

The following compounds were obtained according to similar manners tothose of Examples 1,3,4 and 5.

(1)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)2-(1-tert-butoxycarbonyl-1-methylethoxyimino)acetamido]-3-(3-amino-2,4-dimethyl-1-pyrazolio)methyl-3-cephem-4-c #arboxylate trihydrochloride (syn isomer)

IR (Nu]ol) : 3300, 1780, 1650 cm⁻¹

NMR (D₂ O, δ) : 1.42 (9H, s), 1.47 (6H, s), 1.93 (3H, s), 3.32 (2H,broad s), 3.67 (3H, s), 5.18 (2H, broad s), 5.22 (1H, d, J=5Hz), 5.90(1H, dd, J=8Hz, 5Hz), 7.90 (1H, s), 9.45 (1H, d, J=8Hz)

(2)7β-[2-(5-Amino-1,2,4-thiadiazol-3-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-(3-amino-2,4-dimethyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3320, 3180, 1760, 1650, 1595 cm⁻¹

(3) 7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methyethoxyimino)acetimido]-3-(2-methyl-formamido-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3320, 3180, 1760, 1650, 1595 cm⁻¹

NMR (DMSO-d₆, δ) : 1.46 (6H, s), 3.05-3.37 (2H, m), 3.91 (3H, s),4.90-5.57 (2H, m), 5.06 (1H, d, J=5Hz), 5.71 (1H, dd, J=5, 8Hz), 6.91(1H, d, J=3Hz), 8.02-8.27 (2H, br s), 8.34 (1H, d, J=3Hz), 8.56 (1H, s),9.46 (1H, d, J=8Hz)

(4)7β[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-]3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4#carboxylate(syn isomer)

IR (Nujol) : 3300, 1765, 1640 cm⁻¹

NMR (D₂ O, δ) : 1.58 (6H, s), 3.10 and 3.43 (2H, ABq, J=18Hz), 3.78-3.97(2H, m), 4.26-4.46 (2H, m), 5.15 (2H, br s), 5.26 (1H, d, J=5Hz), 5.87(1H, d, J=5Hz), 5.97 (1H, d, J=3Hz), 7.89 (1H, d, J=3Hz)

(5)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3300, 1775, 1670 cm⁻¹

NMR (D₂ O, δ) : 1.56 (6H, s), 2.31 (3H, s), 3.20 and 3.50 (2H, ABq,J=18Hz), 3.93 (3H, s), 5.23 and 5.47 (2H, ABq, J=15Hz), 5.26 (1H, d,J=5Hz), 5.88 (1H, d, J=5Hz), 6.88 (1H, d, J=3Hz), 8.19 (1H, d, J=3Hz)

(6) 7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(l-carboxy-1-methylethoxyimino)acetamido]-3-(2-methyl-3-ureido-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3300 (broad s), 1770, 1680, 1570 cm⁻¹

EXAMPLE 7

Trifluoroacetic acid (7 ml) was added dropwise to a suspension of7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-1-methylethoxyimino)acetamido]-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carb#oxylate trihydrochloride (3.3 g) and anisole (3.5 ml) in methylenechloride (10 ml) at ambient temperature. After being stirred at the sametemperature for 4 hours, the mixture was poured into diisopropyl ether(300 ml), and the resulting precipitate was collected by filtration. Theprecipitate was dissolved in water (100 ml), and the solution wasadjusted to pH 2 with 5% aqueous solution of sodium bicarbonate. Theaqueous solution was subjected to column chromatography on macroporousnon-ionic adsorption resin "Diaion HP-20". The desired product waseluted with 5% aqueous isopropyl alcohol solution and lyophilized togive7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(l-carboxy-l-methylethoxy-imino)acetamido]-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-ca#rboxylate (syn isomer) (515 mg).

IR (Nujol) : 3325, 1770, 1650, 1630, 1590 cm⁻¹

NMR (D₂ O, δ) : 1.52 (6H, s), 3.19 and 3.37 (2H, ABq, J=18Hz), 3.66 (3H,s), 4.97 and 5.25(2H, ABq, J=15Hz), 5.20 (1H, d, J=5Hz), 5.84 (1H, d,J=5Hz), 5.91 (1H, d, J=3Hz), 7.82 (1H, d, J=3Hz)

EXAMPLE 8

The following compounds were obtained according to a similar manner tothat of Example 7.

(1)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-(3-amino-2,4-dimethyl-l-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3320, 3180, 1760, 1650, 1595 cm⁻¹

NMR (D₂ O, δ) : 1.60 (6H, s), 1.96 (3H, s), 3.10 and 3.37 (2H, ABq,J=18Hz), 3.68 (3H, s), 4.92 and 5.23 (2H, ABq, J=15Hz), 5.22 (1H, d,J=5Hz), 5.86 (1H, d, J=5Hz), 7.68 (1H, s)

(2)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-(4-formamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3300, 1765, 1665, 1605 cm⁻¹

(3)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-(3-formamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3300, 1760, 1660, 1580 cm⁻¹

(4)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-(3-formamido-2,4-dimethyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3300, 1765, 1660, 1600 cm⁻¹

(5)7β-[2-(5-Amino-1,2,4-thiadazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3300, 1760, 1660, 1590 cm⁻¹

(6)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-(3-amino-2,4-dimethyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3350, 1770, 1660, 1600 cm⁻¹

(7)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(l-carboxy-1-methylethoxyimino)acetamido]-3-(2-methyl-3-formamido-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3200-3300, 1760, 1580 cm⁻¹

(8)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(l-carboxy-1-methylethoxyimino)acetamido]-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carb#boxylate (syn isomer) IR : (Nujol) : 3300, 1765, 1640 cm⁻¹

(9)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(l-carboxy-1-methylethoxyimino)acetamido]-3-(3-acetamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3300, 1775, 1670 cm⁻¹

(10)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-(2-methyl-3-ureido-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3300 (broad s), 1770, 1680, 1570 cm⁻¹

PREPARATION 10

To a solution of malononitrile (300 g) and acetic acid (54.6 g) inacetonitrile (2.1 l) was added sodium nitrite (313 g) at 60-65° C. Afterbeing stirred at the same temperature for an hour, ethyl2-bromo-2-methylpropanoate (797 g) was added to the mixture at 65° C.After being refluxed for five hours, the mixture was cooled to theambient temperature and allowed to stand overnight. The resultingmixture was poured into a mixture of water (4.2 ) and diisopropyl ether(4.2 l). The separated organic layer was washed with water and brine,dried over magnesium sulfate and evaporated under reduced pressure togive 2-(1-ethoxycarbonyl-1-methylethoxyimino)propanedinitrile (860 g).

IR (Neat) : 3000, 2250, 1750 cm⁻¹

NMR (CDCl₃, δ) : 1.30 (t, J=7Hz, 3H), 1.69 (s, 6H), 4.25 (q, J=7Hz, 2H)

PREPARATION 11

To a solution of ammonium acetate (1258 g) in methanol (8.54 l) wereadded 2-(1-ethoxycarbonyl-1-methylethoxyimino)propanedinitrile (854 g)and 28% ammonia water (568 ml). After the mixture was stirred at 20° C.for 15 hours, methanol was evaporated under reduced pressure. Theresidue was dissolved in a mixture of water (8 l) and tetrahydrofuran (8l). After the aqueous layer was saturated with sodium chloride, theorganic layer was separated and dried over magnesium sulfate. Aceticacid (368 g) was added to the solution, then the mixture was evaporatedunder reduced pressure, and the residue was triturated with diisopropylether (8 l) to give2-cyano-2-(1-ethoxycarbonyl-1-methylethoxyimino)acetamidine acetate(689.1 g).

IR (Nujol) : 1750, 1670 cm⁻¹

NMR (CDCl₃,δ) : 1.26 (t, J=7Hz, 3H), 1.67 (s, 6H), 2.00 (s, 3H), 4.23(q, J=7Hz, 2H), 8.73 (broad s, 2H)

PREPARATION 12

To a solution of2-cyano-2-(1-ethoxycarbonyl-1-methylethoxyimino)acetamidine acetate (685g) in methanol (6.85 l) was added triethylamine (557 g) at -13˜-15° C.over a 20-minute period and then bromine (344 g) was added thereto atthe same temperature over the same period. The solution was stirred at-13˜-15° C. for 15 minutes, then N,N-dimethylformamide (685 ml) wasadded to the solution at the same temperature, and a solution ofpotassium thiocyanate (209 g) in methanol (2.09 l) was added thereto at-13˜-15° C. over a 30-minute period. After being stirred at -5 ˜0° C.for an hour, the solution was poured into a cold water (5 ˜10° C.) andstirred for an hour under ice-cooling. The resulting precipitates werecollected by filtration, washed with cold water and dried to give2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(l-ethoxycarbonyl-1-methylethoxyimino)acetonitrile(390 g).

IR (Nujol) : 3470, 3280, 3140, 1730, 1625, 1540 cm⁻¹

NMR (CDCl₃, δ) : 1.25 (t, J=7Hz, 3H), 1.67 (s, 6H), 4.23 (q, J=7Hz, 2H),7.86 (broad s 2H).

PREPARATION 13

A mixture of2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-ethoxycarbonyl-1-methylethoxyimino)acetonitrile(8.4 g) in 2N sodium hydroxide was stirred at 60-65° C. for 6 hours,adjusted to pH 4.2 with 6N hydrochloric acid under ice-cooling andwashed with ethyl acetate (100 ml). Tetrahydrofuran (120 ml) was addedto the aqueous solution, and the solution was adjusted to pH 1.0 with 6Nhydrochloric acid under ice-cooling and saturated with sodium chloride.The separated organic layer was evaporated under reduced pressure. Theresidue was dissolved in a saturated aqueous solution (90 ml) of sodiumbicarbonate and activated carbon (200 mg) was added thereto. Afterinsoluble material and activated carbon were removed by filtration, thesolution was adjusted to pH 1.0 with 6N hydrochloric acid underice-cooling. The resulting precipitates were collected by filtration,washed with cold water and dried to give a crude product (6.96 g).

The crude product was recrystallized from propyl alcohol (111 ml) togive(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)aceticacid (4.70 g).

IR (Nujol) : 3430, 3280, 3130, 1740, 1680, 1635, 1540 cm⁻¹

NMR (DMSO-d₆, δ) : 1.46 (s, 6H), 8.23 (broad s, 2H)

PREPARATION 14

A solution of(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-1-methylethoxyimino)aceticacid (17.8 g) and anisole (18 ml) in trifluoroacetic acid (36 ml) wasstirred for 4.0 hours at room temperature. The reaction mixture wasconcentrated under reduced pressure and an aqueous solution of sodiumbicarbonate was added thereto to adjust to pH 6.0. The aqueous solutionwas washed with ethyl acetate, adjusted to pH 3.8 with 6N hydrochloricacid and washed with ethyl acetate. The aqueous layer was separated and6N hydrochloric acid was added thereto to adjust to pH 1.0. After themixture was stirred for 10 minutes, the resultant precipitate wascollected by filtration to give(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)aceticacid (13.0 g).

PREPARATION 15

To a solution of(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)aceticacid (2.74 g) in N,N-dimethylacetamide (45 ml) were addedmethanesulfonyl chloride (1.15 g) and potassium bicarbonate (1.20 g)under cooling in an ice bath. The mixture was stirred for 2.5 hours at5° C. and poured into a cold mixture of water (200 ml), ethyl acetate(200 ml) and 1N hydrochloric acid (6 ml). The mixture was stirred for 5minutes under cooling in an ice bath. The organic layer was separated,washed with cold water (200mlx2) and with a cold saturated aqueoussolution of sodium chloride, dried over magnesium sulfate andevaporated. Toluene was added to the residue. To the mixture was addedmethylene chloride and the mixture was cooled for 10 minutes in an icebath. The crystals were collected by filtration, washed with methylenechloride and air-dried to give methanesulfonyl(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetate(1.1 g).

EXAMPLE 9

A mixture of7β-amino-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylatetrihydrochloride (0.42 g), N-(trimethylsilyl)acetamide (2.5 g) andtetrahydrofuran was stirred for 1.0 hour at room temperature.

Methanesulfonyl(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetate(350 mg) was added thereto at room temperature and stirred for 2.0 hoursat the same temperature. The mixture was poured into diisopropyl ether(50 ml), and the precipitates were collected by filtration, dissolved inwater (50 ml), adjusted to pH 2.0 with an aqueous solution of sodiumbicarbonate and washed with ethyl acetate, and ethyl acetate in theaqueous layer was evaporated. The aqueous layer was subjected to columnchromatography on macroporous non-ionic adsorption resin "Diaion HP-20"and the elution was carried out with 30% aqueous methyl alcoholsolution. Methyl alcohol in the fractions containing the object compoundwas evaporated and the residue was lyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)-acetamido]-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4#-carboxylate(syn isomer)(330 mg).

IR (Nujol) : 3325, 1770, 1650, 1630, 1590 cm⁻¹

NMR (D₂ O, δ) : 1.52 (6H, s), 3.19 and 3.37 (2H, ABq, J=18Hz), 3.66 (3H,s), 4.97 and 5.25 (2H, ABq, J=15Hz), 5.20 (1H, d, J=5Hz), 5.84 (1H, d,J=5Hz), 5.91 (1H, d, J=3Hz), 7.82 (1H, d, J=3Hz).

PREPARATION 16

7β-Amino-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylatetrihydrochloride (5 kg) was dissolved in water (16 l). This solution wassubjected to column chromatography on macroporous non-ionic adsorptionresin "Diaion HP-20".

The desired product was eluted with water. To the eluate (30 l) wasadded acetone (160 l) and the mixture was stirred at room temperaturefor 2 hours. The resulting precipitate was collected by filtration togive7β-amino-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylatehydrochloride dihydrate as crystals (1.802 kg).

IR (Nujol): 3540, 3350, 3150, 1775, 1635, 1585 cm⁻¹

NMR (DMSO-d₆, δ): 3.66 (3H, s), 4.83 (1H, d, J=5Hz), 5.03 (1H, d,J=5Hz), 5.18 and 5.30 (2H, ABq, J=15Hz), 5.85(1H, d, J=3Hz), 7.44 (2H,broad s), 8.08 (1H, d, J=3Hz)

EXAMPLE 10

To N,N-dimethylformamide (231.6 ml) was added 7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer) (38.6 g) at ambient temperature. The mixture was stirred atthe same temperature for 2 hours, and the resultant precipitate wascollected by filtration to give bis(N,N-dimethylformamide) solvate (47.3g) of7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer).

IR (Nujol) : 3280, 3130, 1775, 1670, 1580 cm⁻¹

NMR (D₂ O+NaHCO₃, δ) : 1.53 (6H, s), 2.86 (6H, s). 3.01 (6H, s), 3.10and 3.36 (2H, ABq, J=18Hz), 3.66 (3H, s), 4.96 and 5.23 (2H, ABq,J=15Hz), 5.22 (1H, d, J=5Hz), 5.85 (1H, d, J=5Hz), 5.92 (1H, d, J=3Hz),7.83 (1H, d, J=3Hz), 7.91 (2H, s)

EXAMPLE 11

To a solution of7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer) (0.5 g) in an aqueous sulfuric acid (2M, 1.0 ml) was addedethanol. After the solution was stirred for 1.0 hour, the crystals werecollected by filtration, washed with a solution of water and ethanol(1:5), then washed with ethanol and dried over phosphorus pentoxide togive sulfuric acid salt (480 mg) of7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carb#oxylate (syn isomer).

mp : 194-197° C.

IR (Nujol) : 3320, 3200, 3060, 1770, 1720, 1655, 1590, 1545 cm⁻¹

NMR (D₂ O, δ) : 1.50 (6H, s), 3.33, 3.13 (2H, ABq, J=18Hz), 3.65 (3H,s), 5.20 (1H, d, J=5Hz), 5.22, 4.98 (2H, ABq, J=14Hz), 5.83 (1H, d,J=5Hz), 5.92 (1H, d, J=3Hz), 7.80 (1H, d, J=3Hz)

PREPARATION 17

To a solution of sulfuryl chloride (105.15 g) in methylene chloride(1500 ml) was added a solution of triphenyl phosphite (279 g) inmethylene chloride (300 ml) at -20° C. and the mixture was stirred for30 minutes at -20˜-30° C. To the mixture was added benzhydryl7β-8-(2-hydroxybenzylideneamino)-3-hydroxyethyl-3-cephem-4-carboxylate(300 g) at -20˜-30° C. The mixture was stirred for 30 minutes at-20˜-30° C. and poured into a mixture of ethyl acetate (7.5 l) and 6.5%aqueous potassium carbonate (2.4 l). The organic layer was separated,washed with brine (300 ml) and concentrated in vacuo to 500 ml at 30° C.To the concentrated solution were added N,N-dimethylformamide (300 ml),5-formamido-1-methylpyrazole (187.5 g) and potassium iodide (119.4 g)and the mixture was stirred for 22 hours at 31˜34° C. The reactionmixture was diluted with acetone (600 ml) and the dilute solution wasadded dropwise into the mixture of isopropyl alcohol (6.0 l) anddiisopropyl ether (6.0 l)at 20˜25° C. The resulting precipitate wascollected by filtration and dried in vacuo to give benzhydryl7β-(2-hydroxy-benzylideneamino)-3-(3-formamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylateiodide (514.8 g).

NMR (DMSO-d₆, δ) : 3.60 (2H, br s), 3.75 (3H, s), 5.50 (2H, br s), 5.33(1H, d, J=6Hz), 5.90 (1H, d, J=6Hz), 6.60-7.60 (15H, m), 8.33-8.60 (3H,m), 8.90 (1H, s), 12.00 (1H, m)

IR (Nujol) : 1790, 1725, 1630, 1590, 1230, 1110 cm⁻¹

PREPARATION 18

To a solution of benzhydryl7β-(2-hydroxybenzylideneamino)-3-(3-formamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylateiodide (170 g) in methylene chloride and formic acid was added 35%hydrochloric acid (24.65 g). The mixture was stirred for 3 hours at25˜30° C. and added dropwise into the mixture of acetone (1700 ml) andethyl acetate (3400 ml). The resulting precipitate was collected byfiltration and dried in vacuo to give7β-amino-3-(3-formamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylatehydrochloride (86.7 g).

IR (Nujol) : 1800, 1720, 1650, 1590 cm⁻¹

NMR (DMSO-d₆, δ) : 3.55 (2H, br s), 4.03 (3H, s), 5.30 (2H, br s), 5.33(2H, br s), 6.80-7.60 (2H, m), 8.60 (1H, br s)

PREPARATION 19

To a solution of benzhydryl7β-(2-hydroxybenzylideneamino)-3-(3-formamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylateiodide (510 g) in methylene chloride (1275 ml) and formic acid (1275 ml)was added 35% hydrochloric acid (145 g) at 20˜25° C. The mixture wasstirred for 1 hour at 20˜25° C. and added dropwise into the mixture ofacetone (1020 ml) and ethyl acetate (2040 ml) at the same temperature.The resulting precipitate was collected by filtration and dried in vacuoto give the mixture (255.9 g) of7βamino-3-(3-formamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylatehydrochloride and7β-amino-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylatehydrochloride. The mixture (255.9 g) was dissolved in methanol (1023.6ml). To the solution was added 35% hydrochloric acid (66.6 g) and themixture was stirred for 1 hour at 28 ˜30° C. The insoluble material wasfiltered off and washed with methanol (512 ml). The combined filtratesand washings were added dropwise into the mixture of acetone (2560 ml)and ethyl acetate (5120 ml) at 20˜25° C. The resulting precipitate wascollected by filtration and dried in vacuo to give7β-amino-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylatehydrochloride.

IR (Nujol) : 1800, 1720, 1650, 1600, 1230 cm⁻¹

NMR (DMSO-d₆, δ) : 3.30 and 3.55 (2H, ABq, J=18Hz), 3.70 (3H, s), 5.25(2H, br s), 5.33 (2H, br s), 5.93 (1H, d, J=3Hz), 8.25 (1H, d, J=3Hz)

PREPARATION 20

A solution of sodium cyanate (52 g) in water (400 ml) was added dropwiseto a solution of 1-methyl-5-aminopyrazole (19.4 g) in acetic acid (96ml) and water (192 ml), and the mixture was stirred at ambienttemperature for 7 hours. The reaction mixture was poured into a mixtureof water (400 ml) and ethyl acetate (400 ml). The organic layer wasseparated, washed with saturated aqueous sodium chloride, dried overmagnesium sulfate and evaporated to give 1-methyl-5-ureidopyrazole (14.6g).

IR (Nu]ol) 3300 (broad s), 1735, 1600 (broad s)cm⁻¹

NMR (DMSO-d₆, δ) : 3.63, 3.67 (3H, d, J=2Hz), 6.10, 6.22 (1H, dd,J=2Hz), 7.28, 7.35 (1H, d, J=2Hz).

PREPARATION 21

A mixture of acetic anhydride (11.13 ml) and formic acid (5.93 ml) wasstirred at ambient temperature for 30 minutes. To this solution wasadded 5-amino-1-(2-hydroxyethyl)pyrazole (5 g) under ice-cooling, andthe mixture was stirred at 30-40° C. for 1 hour. The reaction mixturewas poured into a mixture of water, tetrahydrofuran and ethyl acetateand adjusted to pH 6 with aqueous sodium bicarbonate. The organic layerwas separated, and the aqueous layer was extracted with a mixture oftetrahydrofuran and ethyl acetate for three times. The organic layerswere combined, dried over magnesium sulfate and evaporated in vacuo togive 5-formamido-l-(2-formyloxyethyl)pyrazole (5.18 g).

IR (Nujol) : 3180, 1705, 1660 cm⁻¹

NMR (DMSO-d₆, δ) : 4.21-4.61 (4H, m), 6.11 and 6.34 (1H, each d, J=3Hz),7.47 (1H, d, J=3Hz), 8.00 (1H, s), 8.33 (1H, s)

PREPARATION 22

The following compounds were obtained according to a similar manner tothat of Preparation 3.

(1) Benzhydryl7β-tert-butoxycarbonylamino-3-(3-acetamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylateiodide

IR (Nujol) : 1780, 1710 cm⁻¹

NMR (DMSO-d₆, δ) : 1.44 (9H, s), 2.25 (3H, s), 3.43 (2H, br s), 3.74(3H, s), 5.16 (1H, d, J=5Hz), 5.38 (2H, br s), 5.63 (1H, dd, J=8, 5Hz),6.93 (1H, d, J=3Hz), 6.94 (1H, s), 7.15-7.55 (10H, m), 7.97 (1H, d,J=8Hz), 8.25 (1H, d, J=3Hz), 11.13 (1H, s)

(2) Benzhydryl7β-tert-butoxycarbonylamino-3-[3-formamido-2-(2-formyloxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylateiodide

IR (Nujol) : 1780, 1720 cm⁻¹

NMR (DMSO-d₆, δ) : 1.49 (9H, s), 3.43 (2H, br s), 4.14-4.38 (2H, m),4.52-4.73 (2H, m), 5.15 (1H, d, J=5Hz), 5.40 (2H, br s), 5.67 (1H, dd,J=5, 8Hz), 6.88 (1H, s), 7.02 (1H, d, J=3Hz), 7.18-7.52 (l0H, m), 7.94(1H, d, J=8Hz), 7.99 (1H, s), 8.27 (1H, d, J=3Hz), 8.51 (1H, br s)

PREPARATION 23

The following compounds were obtained according to a similar manner tothat of Preparation 5.

(1) Di(trifluoroacetic acid) salt of7β-amino-3-[3-formamido-2-(2-formyloxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate

IR (Nujol) : 1780, 1715, 1660 cm⁻¹

NMR (DMSO-d₆, δ) : 3.53 (2H, br s), 4.28-4.56 (2H, m), 4.78-4.99 (2H,m), 5.29 (2H, br s), 5.53 (2H, br s), 7.14 (1H, d, J=3Hz), 8.22 (1H, s),8.46 (1H, d, J=3Hz), 8.63 (1H, s)

(2) Di(trifluoroacetic acid) salt of7β-amino-3-(3-acetamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate

IR (Nujol) : 1780, 1670 cm⁻¹

NMR (DMSO-d₆, δ) : 2.24 (3H, s), 3.47 (2H, br s), 3.93 (3H, s), 5.22(2H, s), 5.50 (2H, br s), 6.98 (1H, d, J=3Hz), 8.35 (1H, d, J=3Hz)

PREPARATION 24

The following compound was obtained by reacting di(trifluoroacetic acid)salt of7β-amino-3-[3-formamido-2-(2-formyloxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylateaccording to a similar manner to that of Preparation 7.

7β-Amino-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]-methyl-3-cephem-4-carboxylatetrihydrochloride

IR (Nujol) : 3250, 1770, 1700, 1625 cm⁻¹

NMR (DMSO-d₆, δ) : 3.43 (2H, br s), 3.52-3.88 (2H, m), 4.18-4.48 (2H,m), 5.28 (2H, br s), 5.37 (2H, br s), 5.97 (1H, d, J=3Hz), 8.18 (1H, d,J=3Hz)

PREPARATION 25

To a solution of ammonium acetate (73.7 g) in methanol (250 ml) wereadded 2-(1-ethoxycarbonyl-1-methylethoxyimino)propanedinitrile (50.0 g)and 28% ammonia water (33.3 ml) at room temperature. After stirring for15 hours at 20° C., potassium carbonate (33.0 g) was added to thesolution, and the solution was evaporated under reduced pressure. Amixture of water (130 ml) and methylene chloride (110 ml) was added intothe residue. The mixture was adjusted to pH 8.0-8.2 and extracted withmethylene chloride twice. Phthalic acid (35.7 g) was added to theextract at 20-30° C., and then diisopropyl ether (200 ml) was addedthereto. After stirring for 2 hours at the same temperature, theresulting precipitate was collected by filtration, washed withdiisopropyl ether (40 ml) and dried under reduced pressure to givephthalic acid salt (61.4 g) of2-cyano-2-(1-ethoxycarbonyl-1-methylethoxyimino)acetamidine.

mp : 156-158° C.

IR (Nujol) : 3380, 1730, 1700 cm⁻¹

NMR (DMSO-d₆, δ) : 1.20 (3H, t, J=7Hz), 1.66 (6H, s), 4.17 (2H, q,J=7Hz), 7.4-7.7 (2H, m), 7.9-8.2 (2H, m), 12.0 (5H, broad s)

PREPARATION 26

To a suspension of phthalic acid salt (10.0 g) of2-cyano-2-(1-ethoxycarbonyl-1-methylethoxyimino)-acetamidine in methanol(60 ml) were added dropwise triethylamine (9.01 g) and bromine (3.67 g)at -15˜-10° C. After stirring for 15 minutes at the same temperature,N,N-dimethylformamide (6.0 ml) and a solution of potassium thiocyanate(2.23 g) in methanol (22.3 ml) were added dropwise thereto at -15˜-10°C. After stirring for one hour at -5˜0° C., the solution was poured intocold water (300 ml) and the mixture was stirred for one hour underice-cooling. The resulting precipitate was collected by filtration,washed with cold water (60 ml) and dried under reduced pressure to give2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-ethoxycarbonyl-1-methylethoxyimino)-acetonitrile(3.82 g).

IR (Nujol) : 3470, 3280, 3140, 1730, 1625, 1540 cm⁻¹

EXAMPLE 12

1-Methyl-5-ureidopyrazole (1.4 g) was added to a solutionof7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-chloromethyl-3-cephem-4-carboxylicacid trifluoroacetate(syn isomer) (1.24 g) in N,N-dimethylformamide (13ml) and the mixture was stirred at ambient temperature for 4 hours. Thereaction mixture was poured into ethyl acetate (100 ml). Theprecipitates were collected by filtration and successively washed withethyl acetate and diisopropyl ether, and the solid was dissolved inwater (30 ml) and adjusted to pH 2.0 with 10% hydrochloric acid. Thesolution was subjected to column chlomatography on macroporous non-ionicadsorption resin "Diaion HP-20"and eluted with 30% aqueous solution ofmethyl alcohol. The fractions containing the ob]ect compound werecollected, concentrated in vacuo and lyophilized to give7β-[2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-(2-methyl-3-ureido-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer) (0.12 g).

IR (Nujol) : 3300 (broad s), 1770, 1680, 1570 cm⁻¹

NMR (D₂ O, δ) : 1.53 (6H, s), 3.14, 3.44 (2H, ABq, J=18Hz), 3.70 (3H,s), 5.22 (1H, d, J=5Hz), 5.27 (2H, broad s), 5.85 (1H, d, J=5Hz), 6.75(1H, d, J=3Hz), 8.10 (1H, d, J=3Hz)

EXAMPLE 13

The following compounds were obtained according to a similar manner tothat of Example 12.

(1)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-carboxy-methoxyiminoacetamido]-3-(4-formamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3300, 1765, 1665, 1605 cm⁻¹

(2)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-(3-formamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3300, 1760, 1660, 1580 cm⁻¹

(3)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-carboxymethoxyiminoacetamido]-3-(3-formamido-2,4-dimethyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3300, 1765, 1660, 1600 cm⁻¹

(4)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-carboxy-methoxyiminoacetamido]-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3300, 1760, 1660, 1590 cm⁻¹

(5)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-carboxy-methoxyiminoacetamido]-3-(3-amino-2,4-dimethyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3350, 1770, 1660, 1600 cm⁻¹

(6)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-1-methylethoxyimino)acetamido]-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carbox#ylate trihydrochloride (syn isomer)

IR (Nujol) : 3300, 1780, 1720, 1650 cm⁻¹

(7)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-1-methylethoxyimino)acetamido]-3-(3-amino-2,4-dimethyl-1-pyrazolio)methyl-3-cephem-4-ca#rboxylate trihydrochloride (syn isomer)

IR (Nujol) : 3300, 1780, 1650 cm⁻¹

(8)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-(3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3325, 1770, 1650, 1630, 1590 cm⁻¹

(9)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-1-carboxy-1-methylethoxyimino)acetamido]-3-(3-amino-2,4-dimethyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3320, 3180, 1760, 1650, 1595 cm⁻¹

(10)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-(2-methyl-3-formamido-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3200-3300, 1760, 1580 cm⁻¹

(11)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]methyl-3-cephem-4-carboxylate (syn isomer)

IR (Nujol) : 3300, 1765, 1640 cm⁻¹

(12)7β-[2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-(3-acetamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate(syn isomer)

IR (Nujol) : 3300, 1775, 1670 cm⁻¹

What we claim is:
 1. A compound of the formula: ##STR8## wherein R¹ andR⁴ are each amino or a protected amino group, R² is carboxy(lower)alkylor esterifed carboxy(lower)alkyl, andR³ is hydroxy(lower)alkyl or aprotected hydroxy(lower)alkyl,and a pharmaceutically acceptable saltthereof.
 2. A compound of claim 1,wherein R³ is hydroxy(lower)alkyl orlower alkanoyloxy(lower)alkyl.
 3. A compound of claim 2,wherein R² iscarboxy(lower)alkyl or lower alkoxycarbonyl(lower)alkyl.
 4. A compoundof claim 3,wherein R¹ is amino, R³ is hydroxy(lower)alkyl, and R⁴ isamino,
 5. A compound of claim 7,wherein R² is carboxy(lower)alkyl orlower alkoxycarbonyl(lower)alkyl.
 6. A compound of claim 5,wherein R¹ isamino, and R⁴ is amino, lower alkanoylamino or ureido.
 7. A compound ofclaim 1,wherein R² is 1-carboxy-1-methylethyl or carboxymethyl.
 8. Apharmaceutical composition which comprises, as an active ingredient, acompound of claim 1 or a pharmaceutically acceptable salt thereof inadmixture with pharmaceutically acceptable carriers.
 9. A method for thetreatment of infectious diseased which comprises administering acompound of claim 1 or a pharmaceutically acceptable salt thereof tohuman or animals.